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The Protective Mechanism of Experimental Nerve Damage by Baicilin Through Thioredoxin(Trx) Pathway

Author: LiuNan
Tutor: MaJiSheng
School: Jilin Agricultural University
Course: Of Pharmacy
Keywords: Parkinson’s disease baicalin thioredoxin
CLC: R285.5
Type: Master's thesis
Year: 2012
Downloads: 50
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Abstract


Thioredoxin, a small molecule protein with antioxidant activity in vivo, has been widely studied in recent years.It was found that it is closely linked with neurodegenerative disorders such as Alzheimer’s disease and Parkinson’s disease. Scutellaria is a classic Qingrejiedu Chinese Traditional Medicine. Huanglian Jiedu soup, a kind of Traditional Chinese Herbal Compound, has been shown to have a therapeutic effect on Parkinson’s disease clinically.The main ingredient in Huanglian Jiedu soup is scutellaria, and Baicalin is one of the main active components in Scutellaria baicalensis Georgi. Therefore, in this article, we will verify the protective mechanism of baicalin on experimental nerve injury through thioredoxin pathway by a series of experiments in vitro and in vivo, and provide the theoretical basis for the screening and screening of new Chinese medicine throughTrx pathway for Parkinson’s disease.In this experimental, a model of oxidative injury was established by hydrogen peroxide inducing human neuroblastoma cells (SH-SY5Y), and to observe the protective mechanism of Baicalin on experimental nerve cell injury. To detect the cell viability with the Tetrazolium bromide (MTT), and to determine the cell Trx concentration in each group with ELASA, and to test the cell Trx mRNA transcription level in each group with RT-PCR. In vivo, Parkinson’s disease rats were induced by the rotenone to observe the impact of baicalin on rat behavior change, and detecting Trx concentration in rat brain by ELASA. Then to analysis the influence of baicalin on the peroxidation reaction by detecting reactive oxygen species (ROS), glutathione (GSH), malondialdehyde (MDA) level.The primary rat brain microvascular endothelial cells and astrocytes were co-cultured to establish blood brain barrier model in vitro in transwell. The medium containing baicalin was added in donor pool, and collected the medium in different periods, the concentration of Baicain was tested by High Performance Liquid Chromatography in order to analysis the Baicalin permeability and the influence of nerve cells.In vitro, MTT results showed that different concentration of hydrogen peroxide had damage effect on SH-SY5Y cells, and the cell survival rate reduced significantly. The certain concentrational baicalin can significantly enhance SH-SY5Y cells’survival ability which were damaged by the hydrogen peroxide. Baicalin can increase the Trx level in protein level and transcriptional regulation both in vitro and in vivo,and also have protective effect on PD rats behavior change and peroxidation. Results:1. The hydrogen peroxide can induce SH-SY5Y cells oxidative damage, and baicalin can protect these damage cells, strengthen its activity. and this protective effect may be related with the Trx expression. 2. The Parkinson’s disease model established by rats subcutaneous injected rotenone for long time. Baicalin can obviously improve the rats’behavioral changes, and the protection of baicalin might be realized through improving brain peroxide reaction and increasing the Trx expression.3. The rat brain microvascular endothelial cells and astrocytes co-cultured together to establish the blood brain barrier model. The permeability of baicalin should be further researched.

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